• Title/Summary/Keyword: EL4 cells

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Hydrogen Production by the Immobilized Cells of Rhodopseudomonas sp. E15-1 (Rhodopseudomonas sp. E15-1의 균체 고정화에 의한 수소생성)

  • Bae, Moo;Park, Sun-Hee
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.74-80
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    • 1989
  • For improvement of photobiological hydrogen production, Rhodopseudomonas El5-1, a photo-synthetic becterium capable of producing n high yield of hydrogen, was immobilized and conditions for hydrogen production by immobilized cells were examined. The optimum concentration for the combined matrix was obtained when sodium alginate was used at final concentration of 4%. The immobilized cells may reduce the inhibitory effects of nitrogen or oxygen. To minimize the diffusion resistance of the nutrients in alginate gel, the bend size less than 2 mm in diameter was desirable. The immobilized cells were also able to utilize n wide range of organic substrates for the production of hydrogen. The hydrogen producing activity of the immobilized cells was maintained for 20 days without loss of activity during semi-continuous operation of the reactor by feeding of new medium periodically and continuous production of hydrogen could be successfully performed for 30 days.

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Formulation of Ceftriaxone Conjugated Gold Nanoparticles and Their Medical Applications against Extended-Spectrum β-Lactamase Producing Bacteria and Breast Cancer

  • El-Rab, Sanaa M.F. Gad;Halawani, Eman M.;Hassan, Aziza M.
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1563-1572
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    • 2018
  • Gold nanoparticles (AuNP) and their conjugates have been gaining a great deal of recognition in the medical field. Meanwhile, extended-spectrum ${\beta}$-lactamases (ESBL)-producing bacteria are also demonstrating a challenging problem for health care. The aim of this study was the biosynthesis of AuNP using Rosa damascenes petal extract and conjugation of ceftriaxone antibiotic (Cef-AuNP) in inhibiting ESBL-producing bacteria and study of in vitro anticancer activity. Characterization of the synthesized AuNP and Cef-AuNP was studied. ESBL-producing strains, Acinetobacter baumannii ACI1 and Pseudomonas aeruginosa PSE4 were used for testing the efficacy of Cef-AuNP. The cells of MCF-7 breast cancer were treated with previous AuNP and Cef-AuNP at different time intervals. Cytotoxicity effects of apoptosis and its molecular mechanism were evaluated. Ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy established the formation of AuNP and Cef-AuNP. Transmission electron microscope demonstrated that the formed nanoparticles were of different shapes with sizes of 15~35 nm and conjugation was established by a slight increase in size. Minimum inhibitory concentration (MIC) values of Cef-AuNP against tested strains were obtained as 3.6 and $4{\mu}g/ml$, respectively. Cef-AuNP demonstrated a decrease in the MIC of ceftriaxone down to more than 27 folds on the studied strains. The biosynthesized AuNP displayed apoptotic and time-dependent cytotoxic effects in the cells of MCF-7 at a concentration of $0.1{\mu}g/ml$ medium. The Cef-AuNP have low significant effects on MCF-7 cells. These results enhance the conjugating utility in old unresponsive ceftriaxone with AuNP to restore its efficiency against otherwise resistant bacterial pathogens. Additionally, AuNP may be used as an alternative chemotherapeutic treatment of MCF-7 cancer cells.

Harmal Extract Induces Apoptosis of HCT116 Human Colon Cancer Cells, Mediated by Inhibition of Nuclear Factor-κB and Activator Protein-1 Signaling Pathways and Induction of Cytoprotective Genes

  • Elkady, Ayman I;Hussein, Rania A;El-Assouli, Sufian M
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1947-1959
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    • 2016
  • Background: Colorectal cancer (CRC) is a major cause of morbidity and mortality, being the second most common type of cancer worldwide in both men and women. It accounts yearly for approximately 9% of all new cases of cancers. Furthermore, the current chemotherapeutic regimens seem unsatisfactory, so that exploration of novel therapeutic modalities is needed. The present study was undertaken to investigate the inhibitory effects of a crude alkaloid extract (CAERS) of a medicinal herb, Rhazya stricta, on proliferation of CRC HCT116 cells and to elucidate mechanisms of action. To achieve these aims, we utilized MTT, comet, DNA laddering and gene reporter assays, along with Western blot and RT-PCR analyses. Results: We found that CAERS inhibited cell proliferation and induced apoptotic cell death in HCT116 cells. Hallmarks of morphological and biochemical signs of apoptosis were clearly evident. CAERS down-regulated DNA-binding and transcriptional activities of NF-${\kappa}B$ and AP-1 proteins, while up-regulating expression of the Nrf-2 protein. It also down-regulated expression levels of the ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin and VEGF and up-regulated levels of Bax, caspase-3/7 and -9, p53, p21, Nrf-2. Markedly, it promoted mRNA expression levels of cytoprotective genes including the hemeoxygenase-1, NAD(P)H quinine oxidoreductase 1 and UDP-glucuronyltransferase. Conclusions: These findings indicate that CAERS exerts antiproliferative action on CRC cells through induction of apoptotic mechanisms, and suggest CAERS could be a promising agent for studying and developing novel chemotherapeutic agents aimed at novel molecular targets for the treatment of CRC.

Expression of Granulysin and FOXP3 in Cutaneous T Cell Lymphoma and Sézary Syndrome

  • Shareef, Mohamed Moustafa;Elgarhy, Lamia Hamouda;Wasfy, Rania El-Said
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5359-5364
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    • 2015
  • Background: Multiple complex pathways are operable in the evolution of cutaneous T cell lymphomas (CTCLs). These pathways involve interaction between neoplastic T cells and cells of the immune system (especially dendritic cells and the non-malignant T cells). Granulysin is a proinflammatory antimicrobial peptide which has an immune alarmin function, activating dendritic cells, as well as an active role in tumor immunology and prognosis. FOXP3+ regulatory T cells Tregs are an important player in the immune system. Much controversy is found in the literature about the role of Tregs in CTCL. Aim: The present study aimed to investigate the expression of granulysin and FOXP3 in mycosis fungoides (MF), its precursor lesion large plaque parapsoriasis and its leukemic form ;$s\acute{e}ezary$ syndrome (SS). Materials and Methods: Immunohistochemical expression of granulysin and FOXP3 were assessed in lesional skin biopsies taken from 58 patients (4 large plaque parapsoriasis, 48 MF and 6 SS). Results: Granulysin positivity was cytoplasmic and higher in MF than in parapsoriasis en plaque and higher in the more advanced stages of MF (p<0.001). All groups showed significant differences between each other except between MF tumor stage and SS. FOXP3 positivity was nuclear and higher in early stage MF (plaque and patch stages) than in tumor stages and SS (p<0.001). However the FOXP3 count was lower in parapsoriasis en plaque than in other stages of MF. All the groups showed significant differences between each other except between parapsoriasis and SS and between patch and plaque stages of MF. Conclusions: The present study supports a role for granulysin in MF progression and proposes a novel hypothesis about the effect of FOXP3 +veTregs in the suppression of the activity of the neoplastic cells in MF.

Shikonin Exerts Cytotoxic Effects in Human Colon Cancers by Inducing Apoptotic Cell Death via the Endoplasmic Reticulum and Mitochondria-Mediated Pathways

  • Han, Xia;Kang, Kyoung Ah;Piao, Mei Jing;Zhen, Ao Xuan;Hyun, Yu Jae;Kim, Hyun Min;Ryu, Yea Seong;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.27 no.1
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    • pp.41-47
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    • 2019
  • The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated $IC_{50}$ value of $3{\mu}M$ after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G_1$ phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the $PERK/elF2{\alpha}/CHOP$ apoptotic pathway, and mitochondrial $Ca^{2+}$ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER- and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.

A Study on the Mechanical Reliability of Large-area Bi-facial Glass-to-glass Photovoltaic Modules (대면적 양면 태양광 모듈의 기계적 신뢰성 연구)

  • Yohan, Noh;Jangwon, Yoo;Jaehyeong, Lee
    • Current Photovoltaic Research
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    • v.10 no.4
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    • pp.111-115
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    • 2022
  • For the high efficiency of the photovoltaic module, a high-output solar cell, which is the basis of photovoltaic power generation, is required. As the light receiving area of the solar cell increases, the light receiving area of the photovoltaic module also increases. Accordingly, recent trend is to use large-area solar cells such as M6 and M8 instead of M2-based solar cells for manufacturing the photovoltaic module and a study on the mechanical stiffness of the module with increased size is required. In this study, a mechanical load test corresponding to IEC-61215 was performed among the reliability tests of large-area photovoltaic modules. In order to confirm the degree to which the mechanical load test affects the photovoltaic module, the output and EL images were checked by sequentially increasing the pressure by 600 Pa at a pressure of 2400 Pa. Also, factors such as output and efficiency of large-area photovoltaic modules were verified through mechanical load testing of actual large-area photovoltaic modules and the rate of change was very small at 1%.

Production, Isolation, and Purification of L-Asparaginase from Pseudomonas Aeruginosa 50071 Using Solid-state Fermentation

  • El-Bessoumy, Ashraf A.;Sarhan, Mohamed;Mansour, Jehan
    • BMB Reports
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    • v.37 no.4
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    • pp.387-393
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    • 2004
  • The L-asparaginase (E. C. 3. 5. 1. 1) enzyme was purified to homogeneity from Pseudomonas aeruginosa 50071 cells that were grown on solid-state fermentation. Different purification steps (including ammonium sulfate fractionation followed by separation on Sephadex G-100 gel filtration and CM-Sephadex C50) were applied to the crude culture filtrate to obtain a pure enzyme preparation. The enzyme was purified 106-fold and showed a final specific activity of 1900 IU/mg with a 43% yield. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme revealed it was one peptide chain with $M_r$ of 160 kDa. A Lineweaver-Burk analysis showed a $K_m$ value of 0.147 mM and $V_{max}$ of 35.7 IU. The enzyme showed maximum activity at pH 9 when incubated at $37^{\circ}C$ for 30 min. The amino acid composition of the purified enzyme was also determined.

Characterization of Hemolytic Aeromonas sp. MH-8 Responding to the Exposure of Green Tea Catechin, EGCG (녹차 카테킨 EGCG의 노출에 따른 식중독 세균인 용혈성 Aeromonas sp. MH-8의 특성조사)

  • Kim, Dong-Min;Oh, Kye-Heon
    • KSBB Journal
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    • v.31 no.4
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    • pp.228-236
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    • 2016
  • The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.

Effects of irradiation on the mRNA expression of the osteocalcin and osteopontin in MC3T3-E1 osteoblastic cell line (MC3T3-E1 조골세포주의 osteocalcin과 osteopontin mRNA 발현에 미치는 방사선의 영향)

  • Cho Su-Beom;Lee Sang-Rae;Koh Kwang-Joon
    • Imaging Science in Dentistry
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    • v.33 no.3
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    • pp.179-185
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    • 2003
  • Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, particularly on the expression of osteocalcin and osteopontin. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1,4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. After the specimens were harvested, RNA was extracted on the 3rd, 7th, 14th, and 21st day after irradiation. The RNA strands were reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR. Results: The irradiated cells demonstrated a dose-dependent increase in osteocalcin and a dose-dependent decrease in osteopontin mRNA expression compared with the non-irradiated control group, The amount of osteocalcin mRNA expression decreased significantly at the 3rd day after irradiation of 0,5, 1,4, and 8 Gy, and also decreased significantly at the 3rd, 14th, and 21 st day after irradiation in the 8 Gy exposed group compared with the control group, The degree of osteopontin mRNA expression increased significantly at the 7th day after irradiation of 0,5, 1,4, and 8Gy, Conclusion: These results showed that each single dose of 0,5, 1, 4, and 8 Gy influenced the mRNA expression of osteocalcin and osteopontin associated with the calcification stage of osteoblastic cells, suggesting that each single dose affected bone formation at the cell level.

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Involvement of F-Actin Cytoskeleton for Microvilli Formation of Jurkat T Lymphocyte (F-actin cytoskeleton이 Jurkat T 림파구의 microvilli 형성에 미치는 영향)

  • Lee, Jae-Seol;Kim, Hae-Young;Son, Gi-Ae;Kim, Ji-Eun;Moon, Kyoung-Mi;Kim, Kwang-Hyeon;Choi, Woo-Bong;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1401-1406
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    • 2011
  • Morphological changes in immune cells occur due to pathogen infection and natural circulation. T cells produce uropod, filopodia, lamellipodia, and microvilli for inflammation, immunosurvelliance, migration, and diapedesis. Short finger-like microvilli cover the surfaces of circulating mammalian immune cells. The surface features of monocytes and neutrophils are quite different, containing membrane ruffles as their predominant structure. In this study, we present the involvement of actin cytoskeleton regarding T lymphocyte microvilli. From analysis of scanning electron micrographs, Jurkat T lymphocyte microvilli was observed to rapidly disassemble when exposed to the actin-sequestering molecule, cytochalasin D. In contrast to cytochalasin D treatment, we found that median microvillar thickness was enlarged on Jurkat T lymphocytes treated with PMA via Lin-11, Isl-1, Mec-3 Kinase (LIMK) and cofilin signaling. In addition, actin cytoskeleton was involved in polarity formation in EL4 T lymphocytes. These results suggest that microvilli formation or polarity of T lymphocytes are involved in actin cytoskeleton dynamics.